Method of mechanical fractionation of gels and apparatus for performing the same



V. MAIZEL. JR

June 24, 1969 METHOD OF MECHANICAL FRACTIONATION OF GELS AND APPARATUSSheet of3 FOR PERFORMING THE SAME Filed June 17, 1966 INVENTOR. JacobMuizel 3,451,629 METHOD OF MECHANICAL FRACTIONATION OF GELS ANDAPPARATUS Sheet of 3 June 24, 1969 J. v. MAIZEL. JR

FOR PERFORMING THE SAME Filed June 17, 1966 JNVENTOR. Jacob \L Maize!June 24, 1969 J. v. MAIZEL. JR=

METHOD OF MECHANICAL FRACTIONATION OF GELS AND APPARATUS FOR PERFORMINGTHE SAME Sheet Filed June 17. 1966 FIG.7

FRAcT'toN NUMBER.

WPQrIE NWQ mPZDOU FIG.8

mm mm 60 40 so so FRACTION NUMBER INVENTOR. Jacob Maize! (7 L-d/( g 2AATTORNEY FIG.9

United States Patent US. Cl. 2411 Claims ABSTRACT OF THE DISCLOSURE Amethod of mechanical fractionation of gels is shown which includes thesteps of applying pressure to the gel while impinging the same upon apressure responsive element in the direction of the pressure so as todivide the gel into fractions and applying a pressurized liquid to thepressure responsive area so as to carry away the fractions.

A mechanical fractionator for a gel is disclosed which includes aholding container to support the unfractioned gel, a plunger within thecontainer adapted to apply pressure upon the gel, a measuring mechanismfor the pres sure to be applied which includes a drive screw, a motorfor driving the drive screw, a pressure responsive area adapted toreceive the fractioned portions of the gel, and a conduit impinging uponthe pressure responsive area adapted to contain a pressurized fluid soas to carry away the fractioned portions of the gel.

This invention relates to a method of mechanical fractionation of gelsand one form of mechanical fractionator which can perform the same.

Substances distributed in gels including separated proteins, gels, inorder to be properly studied, should be fractioned. Conventionally thisha been done by various methods. The simplest is to hand section thegel, This, of course, is clumsy and bothersome. Electrophoresis has alsobeen used to process the gel. This is expensive, requires highlysophisticated apparatus and is difficult to perform properly.

I have discovered a simple and inexpensive method to fraction gelsinvolving the steps of applying pressure to the gel from one directionwhile impinging the gel upon an area responsive to the direction of thepressure and thence applying pressure fluid to the pressure responsivearea so as to carry away the fractions. These fractions then can beplaced into collecting containers and studied.

The above method can be most conveniently performed by the use ofapparatus also described in the instant specification. The apparatus maybe most broadly described as comprising a holding container adapted tosupport the gel, a pressure responsive area operatively connected withthe container, a pressure producing mechanism selectively actuatableupon the gel disposed within the container, and carrier meansoperatively connected with the pressure re sponsive area.

The above constitutes a brief description of the instant invention andsome of the objects and advantages thereof. Other objects and advantagesof this invention will become obvious to the reader of thisspecification as the description proceeds.

The invention will be further described by reference to the accompanyingdrawings which are made a part of this specification.

FIG. 1 is a top view of the preferred form of apparatus for performingthe mechanical fractionation of gels. In this view the apparatus is inhorizontal position.

FIG. 2 is a side view of the apparatus shown in FIG. 1.

FIG. 3 is a fragmentary detail view of the apparatus shown in FIGS. 1and 2.

FIG. 4 is a detail sectional view, on an enlarged scale of the pressureresponsive area, the container and the pressure producing means and thecarrier means of this invention.

FIG. 5 is a detail view partly in section, of the form of apparatusutilizing pressure fluid as the carrier means and also showing themanner in which the discharged gel fractions are collected, In this viewthe apparatus is in an inverted horizontal position.

FIG. 6 is a view identical to FIG. 5 but showing the gel fractions ledinto a different container.

FIG. 7 is a graph showing the results obtained by the use of thefractioning method of this invention.

FIG. 8 is a graph showing the results obtained by the conventionalelectrophoresis technique.

FIG. 9 is an enlarged detail cross-sectional view of a gel showing thevarious areas as they appear therein after staining.

The instant invention will now be further described by reference to thespecific form thereof shown in the accompanying drawings. In thisconnection, however, the reader is cautioned to note that the specificform of the instant invention as set forth in the specification hereinis for illustrative purposes and for purposes of example only, Variouschanges and modifications may obviously be made within the spirit andscope of this invention and would occur to those skilled in this art.

The mechanical fractionater 11 of this invention is formed with aholding container 12, a pressure responsive area 13 is operativelyconnected with container 12 and consists of a supporting member 14, aconduit (preferably vertical) 15 and a needle valve 16 disposed withinconduit 15. Needle valve 16 comprises plunger 17, knob 18 and threadedstem 19 which in turn meshes with an additional threaded recess 20provided within block 14.

Support member 21 is also preferably provided which is formed with athreaded stem portion 22 which in turn meshes within an additionalthreaded recess 23 provided within block 14. Portion 21 surroundscontainer 12 and, in order to prevent leakage, an O-ring seal 24 isprovided.

An additional conduit 25 (preferably at right angles to conduit 15) isalso provided Within block 14. Conduit 25 is joined to tubes 26 and 27.Tube 26 in turn passes through pump 28 and is joined to tube 29 which isdisposed within a liquid (preferably water) 30 which is carried by flask31. Conduit 27 may selectively be made to impinge upon collectingvessels 45, 46, 47, 48 and 49.

The gel to be fractioned 33 is formed into a plurality of stained areas34 (FIGS. 5, 6 and 9).

The pressure producing mechanism preferably utilized in connection withthis invention comprises a piston 35 which is adapted to move downwardlywithin container 12, The term downwardly relates to the direction ofpiston movement toward valve 16. Piston 35 is secured to drive block 36which in turn moves downwardly on trunnions 37 and 38. Drive block 36includes a threaded opening (not shown) through which drive screw 39passes. Drive screw 39 has a measured thread thereupon and constitutes ameasuring means for the movement of piston 35. Drive block 36 may beclamped into position by means of thumbscrew 40 which is provided with astem 41 which in turn bears against member 39 through an opening withinblock 36.

Screw 39 is driven by reduction gear 42 which in turn is connected togear 43 which is driven by motor 44. It is noted that piston 35 impingesupon an end portion of gel 33 and the opposite end portion of gel 33impinges upon the pressure responsive area. Thus as piston 35 movesdownwardly and the gel is fractioned the sequential integrity of the gelis maintained. That is, even though gel 33 is fractioned the sequence ofthe areas 34 within the unfractioned gel also appears in the fractionedmaterial and each fraction is produced in the same sequence. This isvery important for proper study of the gel material.

With the foregoing specific description, the operation of this inventionmay now be explained.

The gel to be mechanically fractioned 33 is placed within container 12and piston 35 placed in a position above the gel, The piston is adjustedso that the lowermost portion of the gel impinges the uppermost portionof stem 17 (FIG, 6). The drive block 36 is then clamped into positionand motor 44 actauted so that piston 35 is forced downwardly throughcontainer 12 and thereby applies a pressure to the gel 33 to bemechanically fractionated. At the same time, pump 28 is actuated forcingfluid 30 through conduits 29, 26 and 27 causing the pressure fiuid topass through the pressure responsive area and thus carry the crushed gelfractions into conduit 27 which in turn is placed into impingement withthe collecting containers.

As seen by the showings in FIGS. 7 and 8, the result obtained by themethod of this invention and by the use of the apparatus set forththerein are comparable to those obtained by conventional techniques.

The foregoing sets forth the manner in which the objects of thisinvention are achieved.

I claim:

1. A mechanical fractionator for a gel material comprising a holdingcontainer adapted to support said unfractioned gel material, a pressureresponsive area operatively connected with said container and adapted toreceive fractioned portions of said gel, a pressure producing mechanismselectively actuatable upon the gel disposed within said container todivide said gel into fractions, and carrier means including apressurized liquid operatively connected with said pressure responsivearea adapted to carry away the fractioned portions of said gel.

2. A mechanical fractionator as described in claim 1 wherein saidpressure producing mechanism comprising a piston movable within saidholding container and drive means operatively connected with saidpiston,

3. A mechanical fractionator as described in claim 1 wherein saidcarrier means comprise a first conduit in impinging relationship withsaid pressure responsive area, said conduit carrying said pressureliquid.

4. A mechanical fractionator as described in claim 3 wherein saidpressure producing mechanism comprise a piston movable Within said gelcontainer in compressing relationship and motor means operativelyconnected with said piston to drive the same.

5. A mechanical fractionator as described in claim 4 including measuringmeans controlling the movement of said piston.

6. A mechanical fractionator as described in claim 5 wherein saidpressure responsive area comprises a support, a substantially verticalsecond conduit within said support operatively connected with saidholding container, said pressure liquid conduit being horizontallydisposed and connected with said second conduit, and a needle valvewithin said support.

7. A mechanical fractionator as described in claim 6 said measuringmeans comprising a drive screw having measured threads thereupon.

8. A mechnical fractionator for a gel material comprising a holdingcontainer adapted to support unfractioned gel material havingsequentially joined ditfering areas thereupon. a pressure responsivearea operatively connected with said container and in impingement withan end portion of said gel, a pressure producing mechanism selectivelyactuatable upon the opposite end portion of the gel disposed within saidcontainer, said pressure producing mechanism and pressure responsivearea adapted to fraction said gel but to maintain the sequentialintegrity thereof, and carrier means operatively connected with saidpressure responsive area.

9. A method of mechanically fractionating a gel comprising the steps ofapplying pressure to the gel from one direction while simultaneouslyimpinging said gel upon a pressure responsive element so as to dividesaid gel into fractions and applying a pressurized liquid to saidpressure responsive area so as to carry away said fractions.

10. A method of mechanically fractionating a gel formed with a pluralityof diifering areas within said gel but without destroying the sequentialintegrity thereof comprising the steps of placing said gel within aholding element and applying pressure to the gel from one directionwhile simultaneously impinging said gel upon a pressure responsiveelement so as to divide said gel into fractions, and thence applyingcarrying means to said pressure responsive area.

References Cited UNITED STATES PATENTS 2,928,614 3/1960 Emanuel et al.241-39 3,165,266 1/1965 Blum et al 241-1 3,309,032 3/1967 Filz et al241-1 XR FRANK T. YOST, Primary Examiner.

U.S. Cl. X.R. 241-15, 38, 301

